Alberta liesbeth Paul

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Questions related from Alberta liesbeth Paul

What could lead to quenching of Alexa fluor 647 signal in multi-color flow?

Does anybody have experience in complete lost of AF647 signal in multicolor flow? any biochemical explanation? I use a 3 laser (blue; red and violet (488; 635; 405 nm) In particular I am talking...

31 March 2019 8,092 0 View

How to treat/enzyme digest human tonsils for isolation leucocytes?

We want to phenotype all leucocyte populations in human tonsils by multi-color flow cytometry. Do we need an enzymatic treatment for getting out all the leucocytes (DCs, Plasmacells etc.)? If so...

07 September 2018 6,320 2 View

Anybody knows if an antibody against Human ST2/IL-33R that is fluorchrome- conjugated is available ?

I want to determine expression on different Human PBMC populations, cannot find it through my search on the internet. I only found a polyclonal non-conjugated anti-human ST2 from R&D. Or does...

30 October 2017 2,742 5 View

What source of serum is used for human autologous T and B cell cocultures for assessing antigen-specific follicular helper T cell function?

We will assess viral-antigen function in PBMCs after virus challenge. There is discussion of adding human AB serum instead of FBS or autologous serum. Any comments, taking in mind factors in...

29 June 2017 5,106 2 View

What is the experience of using Sepmate tubes for human PBMCs isolation?

We want to change from using Leucosep tube with Ficoll to Sepmate tubes from Stemcell technologies with Lymphoprep which would save us time of isolation. Any comments on differences in recovery,...

29 June 2017 7,181 2 View

How to generate/exxpand follicular helper T cells from human PB?

I need a positive control for my Bcl-6 staining and human Peripheral Blood CXCR5+ T helper cells are negative for this marker according to the literature. I need to make sure the staining works...

10 May 2017 9,132 0 View

How to expand Tfh from human PBMCs besides antiCD3/CD28 stim with ICOSL?

Other cytokines or neutralizing antibodies to cytokines needed

31 March 2017 2,608 0 View

What is the experience with intracellular cytokine staining and surface staining on whole human blood ? Is it better than on isolated PBMC?

I am wondering whether to isolate PBMCs or do the staining on whole human blood, what are the advantages and disadvantages and pittfalls of whole blood staining. I will stain for gamma delta T...

16 December 2014 1,358 7 View

At what point do you add Brefeldin A when stimulating regulatory T cells and Gamma Delta T cells for cytokine detection by flow?

I have used this method before with mice T cells for detecting cytokines and added Bref.A immediately after PMA/Ionomyc. but in the literature there is a lot of differences in the time of...

07 February 2014 8,616 23 View

What is the optimal protocol for detecting IL-17A/IL-10 in short-term cultured (from hrs to ON) gamma delta T cells from human PB ?

I want to investigate cytokines produced by human gamma delta T cells by flow cytometry in PBMCs after isolation by microbeads. My stimulus would be PMA and ionomycine followed by inhibition by...

22 May 2013 7,221 3 View

Do cultured and activate human ILC2 express CD45RO

I have a PBMC culture of 1 week with House dust mite. Besides CD4 T cells (my focus) I see that some CD3-CD4-CD8- population have also divided (as determined by Cell Trace Violet dilution)...

01 January 1970 3,529 1 View